Gross and net nitrogen export from leaves of a vegetative C₄ grass

Nitrogen (N) mobilization from mature leaves plays a key role in supplying amino acids to vegetative and reproductive sinks. However, it is unknown if the mobilized N is predominantly sourced by net N-export (a senescence-related process) or other source of N-export from leaves. We used a new approach to partition gross and net N-export from leaf blades at different developmental stages in Cleistogenes squarrosa (a perennial C₄ grass). Net N-export was determined as net loss of leaf N with age, while gross N-export was quantified from isotopic mass balances obtained following 24 h-long ¹⁵N-labeling with nitrate on 10-12 developmentally distinct (mature and senescing) leaves of individual major tillers. Net N-export was apparent only in older leaves (leaf no. > 7, with leaves numbered basipetally from the tip of the tiller and leaf no. 2 the youngest fully-expanded leaf), while gross N-export was largely independent of leaf age category and was ∼8.4 times greater than the net N-export of a tiller. At whole-tiller level, N import compensated 88 ± 14 (SE) % of gross N-export of all mature blades leading to a net N-export of 0.51 ± 0.07 (SE) μg h^-1 tiller^-1. N-import was equivalent to 0.09 ± 0.01 (SE) d^-1 of total leaf N, similar to reported rates of leaf protein turnover. Gross N-export from all mature blades of a tiller was ∼1.9-times the total demand of the immature tissues of the same (vegetative) tiller. Significant N-export is evident in all mature blades, and is not limited to senescence conditions, implying a much shorter mean residence time of leaf N than that calculated from net N-export. Gross N-export contributes not only to the N demand of the immature tissues of the same tiller but also to N supply of other sinks, such as newly formed tillers. N dynamics at tiller level is integrated with that of the remainder of the shoot, thus highlights the importance of integration of leaf-, tiller-, and plant-scale N dynamics.